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9 results for “Mobile genetic elements”
Number of genes per function within mobile genetic elements in Martinez Arbas, Narayanasamy et. al. (2020)
<p>This repository contains a set of tables separated by COG functional categories and the type of mobile genetic element, i.e. phage or plasmid. Each table contains predicted gene functions for each COG category and information on protospacer-containing contigs (PSCCs) and non-PSCCs</p> <p>This repository is related to the work published in Martinez Arbas, Narayanasamy et. al. (2020).</p>
Spatial Mapping and Host Linking of Mobile Genetic Elements in Complex Microbiomes - Visualizing phage infection
<p>We staged infections at four multiplicities of infection (MOI 0, 0.01, 0.1, and 1), and took snapshots every ten minutes over a 40-minute period. We designed FISH probes targeting the non-coding strand of the <em>gp34</em> gene, which encodes a tail fiber protein and quantified cells with 5 or more MGE spots, less than 5 spots, and no spots</p>
Spatial Mapping of Mobile Genetic Elements and their Cognate Hosts in Complex Microbiomes - Identifying the host taxon of a previously undescribed plasmid
<p>We investigated the taxonomic association of an unknown plasmid within a plaque biofilm of a patient diagnosed with stage 3 periodontitis. We combined long- and short- read sequencing to identify a complete plasmid with minimal homology to any sequence in the RefSeq database. The plasmid carried several predicted genes for mobilization and toxin-antitoxin systems. We designed MGE-FISH probes for the plasmid and combined this MGE-FISH stain with an 18-genera HiPR-FISH panel.</p> <p>Images are labeled by collection time such that the laser order for a given field of view (fov) is: 488nm Lambda, 514nm Lambda, 561nm Lambda, 633nm Airyscan, 405nm Lambda. We used Flye (https://github.com/fenderglass/Flye) to assemble the plasmid using long read Nanopore sequencing only and we used OPERA-MS (https://github.com/CSB5/OPERA-MS) to do hybrid assembly with Illumina short reads and Nanopore long reads. The assemblies are in the fasta files and the reads that map to the assemblies are in the fastq files. </p>
Spatial Mapping of Mobile Genetic Elements and their Cognate Hosts in Complex Microbiomes - Combined MGE and taxonomic mapping
<p>We used rRNA-FISH to stain five common oral genera, <em>Veillonella, Streptococcus, Corynebacterium, Lautropia, </em>and <em>Neisseria, </em>each with a different fluorophore, and we used MGE-FISH to stain the <em>termL</em> gene of the active prophage with a sixth fluorophore.</p> <p>We assembled contigs using combined long- and short-read sequencing and identified a highly abundant plasmid. Alignment of this contig to the plasmid database (PLSDB) showed that the plasmid had previously been observed in <em>Prevotella nigrescens</em> (https://www.ncbi.nlm.nih.gov/datasets/genome/GCF_018127865.1/). We selected two genes from the contig with metallo-β-lactamase (MBL) domains as targets for MGE-FISH (https://www.uniprot.org/uniprotkb/V8CNR4/entry, https://www.uniprot.org/uniprotkb/V8CNR9/entry). We stained both putative MBL genes (<em>pMBL</em>) with the same color using MGE-FISH. For taxonomic mapping, we broadened our target panel by employing HIPR-FISH. We selected a target panel of 18 genera that are highly abundant and prevalent in human plaque. We designed a HiPR-FISH spectral encoding using a 5-fluorophore combinatorial barcoding scheme, whereby each fluorophore represents a binary bit, providing 31 possible barcodes (2^5 - 1 = 31). The fluorophore for MGE-FISH was spectrally distinct from those of HiPR-FISH, enabling simultaneous implementation of both methods.</p> <p>Images are labeled by collection time such that the laser order for a given field of view (fov) is: 488nm Lambda, 514nm Lambda, 561nm Lambda, 633nm Airyscan, 405nm Lambda. We used OPERA-MS (https://github.com/CSB5/OPERA-MS) to do hybrid assembly with Illumina short reads and Nanopore long reads. The assemblies are in the fasta files and the reads that map to the assemblies are in the fastq files. </p>
Spatial Mapping and Host Linking of Mobile Genetic Elements in Complex Microbiomes - Mapping MGEs in oral plaque biofilms at high specificity
<p>We stained for the GFP gene in samples that contained mixtures of plaque and GFP-transformed E. coli. We mapped mefE, an AMR gene located on a plasmid and encoding an antibiotic efflux pump, in the plaque metagenomic data of volunteer A but not volunteer B. To test the efficacy of gel embedding and clearing, we used orthogonal FISH probes, designed to not target any sequence in the plaque. We identified a T7-like prophage via metagenomic analysis and developed probes targeting its capsB gene, which encodes the minor capsid protein. We identified a highly prevalent prophage of the class Caudoviricetes with a large terminase gene, termL, and were able to design a large set of FISH probes to stain in three different colors simultaneously. We identified three non-plasmid AMR genes within metagenome assembled genomes: patA, patB, and adeF.</p>
Spatial Mapping and Host Linking of Mobile Genetic Elements in Complex Microbiomes - Optimization of single molecule MGE FISH
<p>We used <em>Escherichia coli </em>transformed with pJKR-H-tetR plasmids encoding an inducible <em>GFP</em> gene as a model system to assess and optimize MGE-FISH on a confocal microscope. We designed FISH probes for the non-coding strand of the <em>GFP</em> gene, used non-transformed <em>E. coli </em>as a negative control, and tested six different FISH protocols.<strong> </strong></p>
Spatial Mapping and Host Linking of Mobile Genetic Elements in Complex Microbiomes - Combined taxonomic mapping and MGE mapping
<p>We used rRNA FISH to stain five common oral genera, <em>Veillonella, Streptococcus, Corynebacterium, Lautropia, </em>and <em>Neisseria</em>, each with a different fluorophore, and we used MGE-FISH to stain the <em>termL</em> gene of an active prophage with a sixth fluorophore. </p> <p>We chose a target panel of 18 genera that are highly abundant and prevalent in human plaque and designed a HiPR-FISH probe panel using a 5-fluorophore combinatorial barcoding scheme. Using MGE-FISH, we stained a plasmid carrying mefE, subunit of a major-facilitator-superfamily antibiotic efflux pump. </p>
Additional data and code for "You can move, but you can't hide: identification of mobile genetic elements with geNomad"
<ul> <li><strong>benchmark_data:</strong> Data used to train and evaluate the classification models.</li> <li><strong>giant_virus_data:</strong> Sequences and metadata of giant viruses identified in public metagenomes.</li> <li><strong>neural_network_training:</strong> Code used to train geNomad's neural network-based classification model.</li> <li><strong>provirus_data:</strong> Data used to train and evaluate the conditional random field model employed by geNomad to identify provirus regions.</li> <li><strong>reference_sequences:</strong> Sequences of chromosomes, plasmids, and viruses that were used to build geNomad's marker dataset and to generate the training data for the classification models.</li> </ul>
Complete genomes of Asgard archaea reveal diverse integrated and mobile genetic elements
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